ABSTRACT
Pa0665 in Pseudomonas aeruginosa shares homologous sequences with that of the essential A-type iron-sulfur (Fe-S) cluster insertion protein ErpA in Escherichia coli. However, its essentiality in P. aeruginosa and its complementation with E. coli erpA has not been experimentally examined. To fulfill this task, we constructed plasmid-based ts-mutant Δpa0665/pTS-pa0665 using a three-step protocol. The mutant displayed growth defects at 42 °C, which were complemented by expressing ec.erpA. Microscopic observations indicated a petite cell phenotype for Δpa0665/pTS-pa0665 at 42 °C, correlated with the downregulation of the oprG gene. RNA sequencing revealed significant transcriptional changes in genes associated with the oxidative phosphorylation (OXPHOS) system, aligning with reduced ATP levels in Δpa0665/pTS-pa0665 under 42 °C. Additionally, the ts-mutant showed heightened sensitivity to H2O2 at 42 °C. Overall, our study demonstrates the essential role of pa0665 for OXPHOS function and is complemented by ec.erpA. We propose that the plasmid-based ts-allele is useful for genetic analysis of essential genes of interest in P. aeruginosa.
Subject(s)
Bacterial Proteins , Oxidative Phosphorylation , Pseudomonas aeruginosa , Pseudomonas aeruginosa/genetics , Pseudomonas aeruginosa/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Mutation , Escherichia coli/genetics , Escherichia coli/metabolism , Iron-Sulfur Proteins/genetics , Iron-Sulfur Proteins/metabolism , Gene Expression Regulation, BacterialABSTRACT
Polycyclic aromatic hydrocarbons (PAHs) are organic pollutants and manufactured substances conferring toxicity to human health. The present study investigated whether pyrene, a type of PAH, harms rats. Our research provides an effective feature selection strategy for the animal dataset from Wenzhou Medical University's Experimental Animal Center to thoroughly examine the impacts of PAH toxicity on rat features. Initially, we devised a high-performance optimization method (SCBA) and added the Sobol sequence, vertical crossover and horizontal crossover mechanisms to the bat algorithm (BA). The SCBA-KELM model, which combines SCBA with the kernel extreme learning machine model (KELM), has excellent accuracy and high stability for selecting features. Benchmark function tests are then used in this research to verify the overall optimization performance of SCBA. In this paper, the feature selection performance of SCBA-KELM is verified using various comparative experiments. According to the results, the features of the genes PXR, CAR, CYP2B1/2 and CYP1A1/2 have the most impact on rats. The SCBA-KELM model's classification performance for the gene dataset was 100%, and the model's precision value for the public dataset was around 96%, as determined by the classification index. In conclusion, the model utilized in this research is anticipated to be a reliable and valuable approach for toxicological classification and assessment.
Subject(s)
Algorithms , Polycyclic Aromatic Hydrocarbons , Humans , Animals , Rats , Pyrenes/toxicity , Polycyclic Aromatic Hydrocarbons/toxicityABSTRACT
BACKGROUND: Nucleolin (NCL, C23) is a multifunctional phosphoprotein that plays a vital role in modulating the survival, proliferationand apoptosis of cancer cells. However, the effects of NCL on cervical cancer and the underlying mechanisms behind this are poorly understood. METHODS: Lentiviral transfection technology was used to construct NCL knockdown cell lines. MTT, colony formation assays, and tumorigenic assays in vivo were performed to observe cell proliferation. HOECHST 33342 staining, flow cytometry, and caspase activity assay were used to test cell apoptosis. RNA-Seq, Western blotting, and RT-PCR were conducted to investigate the specific molecular mechanism. RESULTS: NCL knockdown inhibited cell proliferation and promoted apoptosis both in vivo and in vitro. Mechanistic studies revealed that NCL knockdown inhibited the PI3K/AKT pathway by upregulating FGF, ITGA, TNXB, VEGF, Caspase 3, and Bax, as well as by downregulating AKT, GNB4, CDK6, IL6R, LAMA, PDGFD, PPP2RSA and BCL-2. In addition, the expression levels of apoptosis-related genes after using a PI3K inhibitor LY294002 were consistent with shRNA studies, while treatment with a 740Y-P agonist showed the opposite effect. CONCLUSIONS: Our findings indicate that downregulation of NCL may be a novel treatment strategy forcervical cancer.
Subject(s)
Phosphatidylinositol 3-Kinases , Phosphoproteins , Proto-Oncogene Proteins c-akt , RNA-Binding Proteins , Apoptosis/genetics , Cell Line, Tumor , Cell Proliferation , Down-Regulation , HeLa Cells , Humans , Phosphatidylinositol 3-Kinases/metabolism , Phosphoproteins/metabolism , Proto-Oncogene Proteins c-akt/metabolism , RNA-Binding Proteins/metabolism , Signal Transduction , NucleolinABSTRACT
Gold (Au) can be used as an ideal metal electrocatalyst for ethanol and glucose oxidation reactions due to its high performance-to-cost ratio. In this paper, the Au aerogel with high-index facets was synthesized by using the laser ablation in liquid technology, which can improve the electrocatalytic activity of Au. The as-prepared Au aerogel showed excellent mass activity and specific activity toward ethanol oxidation reaction, which are 4.6 times and 2.1 times higher than Au/C, respectively. The 3D porous nature and rich defect of the Au aerogel provide more active sites. In addition, the high-index facets with under-coordinated atoms enhance the adsorption of ethanol and glucose molecules, thus improving the intrinsic catalytic activity of Au aerogel. The effect of high-index facets has also been investigated by density functional theory calculations. Furthermore, the Au aerogels also show good electrocatalytic activity and stability toward glucose oxidation reaction. These results are conducive to promote the practical application of Au in electrocatalysis.
ABSTRACT
Polycyclic aromatic hydrocarbons (PAHs) are a group of persistent organic global environmental pollutants and cause harmful effects on human health. Here, we evaluated adverse effects of chrysene, which is a four-ring PAH and an important member of 16 priority PAHs, on the liver. Chrysene was detected in some common raw and cooked Chinese food samples. Hepatotoxicity including increased relative liver weight, hepatocyte swelling and degeneration, and elevated serum alanine aminotransferase (ALT) levels were observed in chrysene-exposed C57BL/6 mice. Glutamine treatment effectively ameliorated chrysene-induced mice liver injury by decreasing serum ALT levels. Chrysene induced mice hepatic glutathione depletion and oxidative DNA damage with increased 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. Hepatic expression levels of the aryl hydrocarbon receptor (AhR), AhR-related target genes including CYP1A1, CYP1A2 and CYP1B1, and AhR nuclear translocator (ARNT) were significantly increased in chrysene-exposed C57BL/6 mice. Chrysene induced mice hepatic mRNA levels of the nuclear factor erythroid 2-related factor 2 (Nrf2) and Nrf2-mediated phase II detoxifying and antioxidant enzymes including NQO1, UGT1A1, UGT1A6, SULT1A1, GSTm1, GSTm3, Catalase (CAT), GPx1, and SOD2. We found that chrysene had toxic effects including increased relative liver weight and elevated serum ALT levels on AhR+/+ mice but not AhR-/- mice. Chrysene significantly induced hepatic mRNA levels of CYP1A1 and CYP1A2 in AhR+/+ mice but not AhR-/- mice. To our knowledge, this study is the first to demonstrate that hepatotoxicity causes by chrysene is dependent on AhR, and Nrf2 plays an important regulation role in protection against oxidative liver injury induced by chrysene.
Subject(s)
Chemical and Drug Induced Liver Injury , Polycyclic Aromatic Hydrocarbons , Animals , Chrysenes , Cytochrome P-450 CYP1A1 , Mice , Mice, Inbred C57BL , Receptors, Aryl Hydrocarbon/geneticsABSTRACT
Background: Compared to commonly-used green space indicators from downward-facing satellite imagery, street view-based green space may capture different types of green space and represent how environments are perceived and experienced by people on the ground, which is important to elucidate the underlying mechanisms linking green space and health. Objectives: This study aimed to evaluate machine learning models that can classify the type of vegetation (i.e., tree, low-lying vegetation, grass) from street view images; and to investigate the associations between street green space and socioeconomic (SES) factors, in Los Angeles County, California. Methods: SES variables were obtained from the CalEnviroScreen3.0 dataset. Microsoft Bing Maps images in conjunction with deep learning were used to measure total and types of street view green space, which were compared to normalized difference vegetation index (NDVI) as commonly-used satellite-based green space measure. Generalized linear mixed model was used to examine associations between green space and census tract SES, adjusting for population density and rural/urban status. Results: The accuracy of the deep learning model was high with 92.5% mean intersection over union. NDVI were moderately correlated with total street view-based green space and tree, and weakly correlated with low-lying vegetation and grass. Total and three types of green space showed significant negative associations with neighborhood SES. The percentage of total green space decreased by 2.62 [95% confidence interval (CI): -3.02, -2.21, p < 0.001] with each interquartile range increase in CalEnviroScreen3.0 score. Disadvantaged communities contained approximately 5% less average street green space than other communities. Conclusion: Street view imagery coupled with deep learning approach can accurately and efficiently measure eye-level street green space and distinguish vegetation types. In Los Angeles County, disadvantaged communities had substantively less street green space. Governments and urban planners need to consider the type and visibility of street green space from pedestrian's perspective.
Subject(s)
Parks, Recreational , Residence Characteristics , Humans , Los Angeles , Machine Learning , Poaceae , Socioeconomic Factors , TreesABSTRACT
This work investigates the excitation of dense comb-like enhanced leaky mode resonance (eLMR) in tilted fiber Bragg grating (TFBG) integrated with indium tin oxide (ITO) nanocoating. The ITO overlay leads to a large reduction in mode loss and a great increase of propagation length for s-polarized leaky modes, which means the leaky modes become guided. The guidance of leaky modes enhances significantly the interaction with the core guided mode, which leads to the generation of strong dense comb-like eLMR. The results show that the ultra-narrow eLMR bands present promising sensing performance with an extended measurement range and provide advantages of high Q measurement over the case of surface plasmon resonance (SPR) and lossy mode resonance (LMR). The similarities and differences between the eLMR and SPR and LMR are also discussed. This study offers new opportunities to develop eLMR-based multifunctional fiber-optic devices with high performance.
ABSTRACT
In contrast to reactive inhibition, intentional inhibition is the internally generated decision to "stop" without any external signals. Whether adults and adolescents' neural correlates on these two inhibitions have any differences is still unknown. We measured 20 adults and 21 adolescents' ERP-related N2 using a free-choice Go/Nogo task. The results of the adult's group showed that the mean amplitude and peak latency of intentional Nogo-N2 did not differ from the reactive Nogo-N2. In contrast, the mean amplitude and peak latency for reactive Nogo-N2 in the adolescent group was significantly greater than what was observed for the intentional Nogo-N2. Comparison across groups revealed that the mean amplitude and peak latency of reactive Nogo-N2 were significantly greater in adolescents than in adults, while intentional Nogo-N2 did not differ between groups. These findings may indicate that adolescents lack of self-control is more reflected in reactive inhibition, if adolescents decide whether to do, they will be as good as adults.
Subject(s)
Electroencephalography/methods , Inhibition, Psychological , Reactive Inhibition , Adult , Female , Humans , Male , Young AdultABSTRACT
Polycyclic aromatic hydrocarbons (PAHs) are a class of pervasive global environmental pollutants and adversely affect human health. Among PAHs, phenanthrene and anthracene are isomers consisting of three benzene rings. In the present study, we have made comparisons of constitutive androstane receptor (CAR) activation and toxic effects on the liver between these two isomers. Phenanthrene, but not anthracene, significantly induced promoter activity and gene expression of human drug metabolizing enzyme CYP2B6 in HepG2 cells and human primary hepatocytes, respectively. Phenanthrene, but not anthracene, significantly increased CYP2B10 expression levels and caused hepatotoxicity in mice. Phenanthrene induced the nuclear accumulation of CAR in the liver of wild-type mice, but not CAR-/- mice. Hepatocellular necrosis, elevated expression levels of some CAR-related genes such as CYP2B10, CYP3A11, UGT1A1, SULT2A1 and GSTM3, and lower hepatic glutathione levels were found in phenanthrene-exposed wild-type mice but not CAR-/- mice. Additionally, phenanthrene and anthracene were detected in both raw and grilled lamb samples. The average concentrations of phenanthrene were much higher than those of anthracene in these samples. This study is the first to demonstrate that phenanthrene, but not its isomer anthracene, effectively activates both human and mouse nuclear receptor CAR, and CAR plays a crucial role in phenanthrene-induced mouse hepatotoxicity. Compared with anthracene, K region may be an important electronic structure of phenanthrene for activation of CAR. Dietary consumption of PAHs-contaminated food is an important exposure route for humans. Exposure to phenanthrene may affect human health especially associated with liver.
Subject(s)
Anthracenes/pharmacology , Chemical and Drug Induced Liver Injury/metabolism , Liver/drug effects , Liver/metabolism , Phenanthrenes/pharmacology , Animals , Aryl Hydrocarbon Hydroxylases/metabolism , Cell Line, Tumor , Constitutive Androstane Receptor , Cytochrome P450 Family 2/metabolism , Gene Expression/drug effects , Glucuronosyltransferase/metabolism , Glutathione Transferase/metabolism , Hep G2 Cells , Hepatocytes/drug effects , Hepatocytes/metabolism , Humans , Male , Mice , Mice, Inbred C57BL , Receptors, Cytoplasmic and Nuclear/metabolism , Sulfotransferases/metabolismABSTRACT
Polycyclic aromatic hydrocarbons (PAHs) are a group of persistent organic pollutants primarily formed from the incomplete combustion of carbonaceous materials, and have adverse effects on human health. In this study, we investigated whether pyrene, a PAH consisting of 4 fused benzene rings, has adverse effects on rat. Adult male Sprague-Dawly rats were treated daily by oral gavage with vehicle (corn oil) or pyrene at doses of 375, 750, 1500, or 2200 mg/kg/day for 4 days. The results showed that pyrene caused hepatotoxicity in rats. When compared with the control group, relative liver weights, plasma alanine aminotransferase, and direct bilirubin levels significantly increased after pyrene exposure. Hepatocyte swelling and degeneration and decreased hepatic total glutathione (GSH) levels were also found in pyrene-exposed rats. We further observed that mRNA levels of several hepatic metabolizing enzymes regulated by constitutive androstane receptor (CAR) such as CYP2B1 and CYP2B2 significantly increased in pyrene-exposed rats. These results suggest that decreased GSH levels, elevated hepatic metabolizing enzyme gene expression, and CAR activation are important contributors for pyrene-induced hepatotoxicity in rats. Additionally, we found pyrene significantly induced plasma inflammatory indices including white blood cell and lymphocyte counts. We also observed that pyrene exposure increased relative weight of kidneys and disrupted kidney function with elevated urea and creatinine levels in rats.
Subject(s)
Chemical and Drug Induced Liver Injury/pathology , Hepatocytes/drug effects , Kidney Diseases/chemically induced , Kidney/drug effects , Pyrenes/toxicity , Animals , Constitutive Androstane Receptor , Glutathione/metabolism , Hepatocytes/metabolism , Hepatocytes/pathology , Kidney/metabolism , Kidney/pathology , Kidney Diseases/pathology , Male , Rats , Rats, Sprague-Dawley , Receptors, Cytoplasmic and Nuclear/metabolism , Toxicity TestsABSTRACT
Hepatocellular carcinoma (HCC) is a major cause of cancer-related death worldwide. As vectors for intercellular information exchange, the potential role of extracellular vesicles (EVs) in HCC formation, progression and therapy has been widely investigated. In this review, we explore the current status of the researches in this field. Altogether there is undeniable evidence that EVs play a crucial role in HCC development, metastasis. Moreover, EVs have shown great potential as drug delivery systems (DDSs) for the treatment of HCC. Exosomal miRNAs derived from HCC cells can enhance transformed cell growth in recipient cells by modulating the expression of transforming growth factor-ß activated kinase-1(TAK1) and downstream signaling molecules. Furthermore, vacuolar protein sortin 4 homolog A(VPS4A) and insulin-like growth factor(IGF)-1 regulate exosome-mediated miRNAs transfer. Immune cells- derived EVs containing integrin αMß2 or CD147 may facilitate HCC metastasis. In addition, EVs-mediated shuttle of long non-coding RNAs (lncRNAs), specifically linc- VLDLR and linc-ROR promote chemoresistance of malignant cells. Heat shock proteins (HSPs)-harboring exosomes derived from HCC tumor cells increase the antitumor effect of natural killer (NK) cells, thus enhancing HCC immunotherapy. Indeed, inhibition of HCC tumor growth has been associated with tumor cell-derived exosomes (TEX)-pulsed dentritic cells (DCs). Exosomes are also essential in liver metastasis during colorectal carcinoma (CRC) and pancreatic ductal adenocarcinomas (PDAC). Therefore, as nucleic acid and drug delivery vehicles, EVs show a tremendous potential for effective treatment against HCC.
Subject(s)
Carcinoma, Hepatocellular/metabolism , Carcinoma, Hepatocellular/pathology , Exosomes/metabolism , Extracellular Vesicles/metabolism , Liver Neoplasms/metabolism , Liver Neoplasms/pathology , Animals , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/therapy , Cell-Derived Microparticles/metabolism , Cholangiocarcinoma , Disease Progression , Drug Delivery Systems , Drug Resistance, Neoplasm , Gene Expression Regulation, Neoplastic , Humans , Immunotherapy , Liver Neoplasms/genetics , Liver Neoplasms/therapy , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Neoplasm Metastasis , Signal TransductionABSTRACT
Laboratory animals and animal experiments are foundations and important support conditions for life sciences, especially for medical research. The animal experiments have drawn extensive attention from the society because of the ethical issue. This paper takes Wenzhou Medical University as an example to give a brief introduction to the ethical review about laboratory animals in the university so as to further draw attention and concerns from the public about the ethical issue of laboratory animals. We successively introduce its scientific projects, nurturing environment and ethical review of laboratory animals.
Subject(s)
Animal Experimentation/ethics , Animals, Laboratory , Animals , UniversitiesABSTRACT
The aim of this study was to elucidate the effects of zedoary turmeric oil (ZTO) on P450 activities (CYP1A2, CYP2C9, CYP2C19, CYP2B6, CYP2D6 and CYP3A4) in rats with liver cirrhosis induced by thioacetamide (TAA). For the induction of liver cirrhosis, rats were given TAA in their drinking water at a concentration of 0.03% for consecutive 5 weeks and then 0.04% for the next consecutive 5 weeks throughout the establishment of cirrhosis. Then the cirrhotic rats were ip given saline, ZTO 100, 200 and 400 mg/kg, respectively, once daily for 2 weeks. When cirrhosis model was established at week 10, all rats of five groups were administered intragastrically with 15 mg/kg phenacetin, 0.6 mg/kg tolbutamide, 15 mg/kg omeprazole, 15 mg/kg bupropion, 15 mg/kg metoprolol, and 10 mg/kg midazolam. Blood samples were collected at a series of time-points and the concentrations of probe drugs in plasma were determined by HPLC-MS/MS. The degree of liver cirrhosis was assessed by HE staining. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) from the model group increased by approximately 4-fold, and a decreased level of albumin (Alb) was also observed, as compared to the control group (P < 0.05). However, ZTO was found to reverse those changes of serum levels observed in the model group, and the 200 mg/kg ZTO treatment group showed the most obvious reverse tendency with significantly decreased ALT, AST and increased Alb levels (P < 0.05). The results indicated that ZTO with the dose of 100 mg/kg could inhibit the activities of CYP450 isoforms CYP2C9 and CYP2D6 in vivo in cirrhotic rats induced by TAA, while ZTO with the dose of 400 mg/kg could induce the activity of CYP2C19 in vivo in cirrhotic rats induced by TAA. However, ZTO showed no influence on cirrhotic rat hepatic CYP1A2, CYP2B6 and CYP3A4 activity in vivo. This has certain guiding significance to clinical treatment.
Subject(s)
Curcuma , Cytochrome P-450 Enzyme System/metabolism , Liver Cirrhosis/metabolism , Liver/drug effects , Plant Extracts/pharmacology , Animals , Disease Models, Animal , Liver/metabolism , Liver/pathology , Liver Cirrhosis/chemically induced , Liver Cirrhosis/pathology , Male , Rats , Rats, Sprague-Dawley , ThioacetamideABSTRACT
A sensitive and selective liquid chromatography tandem mass spectrometry method for determination of azasetron hydrochloride in rabbit plasma was developed. After addition of doxapram hydrochloride as internal standard (IS), protein precipitation by 10% trichloroacetic acid was used as sample preparation. Chromatographic separation was achieved on a Zorbax SB-C(18) (2.1 × 50 mm, 3.5 µm) column with acetonitrile-water as mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring mode was used to quantification using target fragment ions m/z 349.9 â 223.5 for azasetron hydrochloride and m/z 378.9 â 291.8 for the IS. Calibration plots were linear over the range of 6-1000 ng/mL for azasetron hydrochloride in plasma. The lower limit of quantitation for azasetron hydrochloride was 6 ng/mL. The mean recovery of azasetron hydrochloride from plasma was in the range 85.6-92.7%. The RSDs of intra-day and inter-day precision were both less than 12%. This method is simple and sensitive enough to be used in pharmacokinetic research for determination of azasetron hydrochloride in rabbit plasma.
Subject(s)
Bridged Bicyclo Compounds, Heterocyclic/blood , Chromatography, Liquid/methods , Oxazines/blood , Tandem Mass Spectrometry/methods , Animals , Bridged Bicyclo Compounds, Heterocyclic/pharmacokinetics , Calibration , Drug Stability , Linear Models , Male , Oxazines/pharmacokinetics , Rabbits , Reproducibility of Results , Sensitivity and Specificity , Spectrometry, Mass, Electrospray IonizationABSTRACT
A sensitive and selective liquid chromatography-tandem mass spectrometry method for the determination of piracetam in rat plasma was developed and validated over the concentration range of 0.1-20 µg/mL. After addition of oxiracetam as internal standard, a simplified protein precipitation with trichloroacetic acid (5%) was employed for the sample preparation. Chromatographic separation was performed by a Zorbax SB-Aq column (150 × 2.1 mm, 3.5 µm). The mobile phase was acetonitrile-1% formic acid in water (10:90 v/v) delivered at a flow rate of 0.3 mL/min. The MS data acquisition was accomplished in multiple reaction monitoring mode with a positive electrospray ionization interface. The lower limit of quantification was 0.1 µg/mL. For inter-day and intra-day tests, the precision (RSD) for the entire validation was less than 9%, and the accuracy was within the 94.6-103.2% range. The developed method was successfully applied to pharmacokinetic studies of piracetam in rats following single oral administration dose of 50 mg/kg.
Subject(s)
Chromatography, Liquid/methods , Piracetam/blood , Tandem Mass Spectrometry/methods , Acetonitriles , Animals , Drug Stability , Linear Models , Male , Piracetam/chemistry , Piracetam/pharmacokinetics , Pyrrolidines/analysis , Pyrrolidines/chemistry , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and Specificity , Spectrometry, Mass, Electrospray Ionization/methodsABSTRACT
Vitellin (Vt) and vitellogenin (Vg) profiles were analyzed in Pteromalus puparum, a pupal endoparasitoid of Pieris rapae. Non-denaturing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses indicated that both native Vt and Vg were likely 370 kDa in size, consisting of two subunits of approximate 206 and 165 kDa. An indirect double antibody enzyme-linked immunosorbent assay (ELISA) for monitoring hemolymph Vg and ovarian Vt levels was developed using a monoclonal antibody and a polyclonal antibody made specially against P. puparum Vt. The synthesis and uptake of Vg in this wasp was initiated immediately after adult eclosion. The hemolymph Vg and ovarian Vt reached their highest level of 0.58 and 4.51 microg per female 24 and 48 h after adult eclosion, respectively. Both Vg synthesis and uptake were in parallel with ovarian development. The Vt levels in the developing embryos decreased progressively except 12h after parasitism. Meanwhile, nine new polypeptides with sizes ranging from 59.2 to 151 kDa, possibly resulting from the limited proteolysis of Vt originally accumulated in newly laid eggs, were detected de-novo during embryonic development using Western blotting with the monoclonal antibody against Vt. These studies provide the basis for future investigation into endocrinal regulations of vitellogenesis and understanding the reproductive strategy in this wasp.